体内电转染增强基因免疫诱导的HBV特异性体液免疫应答

Electroporation In Vivo Enhances the Humoral Immune Re-sponse against HBV-preS2/S following Gene Immunization

为观察体内电转染对HBV基因免疫诱导的特异性体液免疫应答的调节作用,将HBV-preS2/S编码基因插入pVAON33载体构建重组质粒pVAON33-preS2/S,运用肌肉注射法对BALB/c小鼠进行基因免疫(100ug质粒DNA.100ul.只)。以pVAON33-preS2/S、pVAON33-preS2/S(体内电转染)为实验组,并以pVAON33空质粒为对照。按期采集免疫小鼠血清。采用ELISA法检测免疫小鼠血清特异性抗HBs-IgG抗体。结果显示,pVAON33-preS2/S免疫小鼠后可诱导产生特异性抗HBs-IgG抗体,到第7周时其OD值为0.73±0.18(P/N为2.13),而pVAON33-preS2/S(体内电转染)组诱导了更高水平的特异性抗HBs-IgG抗体,第7周时OD值为1.30±0.45(P/N为3.79),两组比较有显著性差异(P<0.05)。本研究表明体内电转染可明显增强HBV基因免疫诱导的体液免疫应答。

To investigate the influence of electroporation in vivo on humoral immune response induced by HBVDNA vaccine, HBV-preS2/S coding sequence was introduced into the eukaryotic expression vector pVAON33and identified by PCR and DNA sequencing analysis. Female BALB/c mice were primed by i.m. gene immu-nization with different recombinant plasmids on day 0, then given electroporation in vivo in one pVAON33-preS2/S group. The levels of specific IgG in sera collected at the indicated times from each group weredetermined by ELISA. HBsAg specific antibody response was elicited in groups primed with plasmids pVAON33-preS2/S and the level of anti-HBs-IgG antibodies was 0.730.18 at 7 weeks after immunization. However, theresponse against HBsAg in the group primed with pVAON33-preS2/S (electroporation in vivo)was significanthigher than that in pVAON33-preS2/S group (p<0.05). These results indicated electroporation in vivo couldenhance humoral immune response against HBV induced by gene immunization.